Please use this identifier to cite or link to this item: https://doi.org/10.1007/s11248-007-9101-3
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dc.titleTransgene integration and organization in Cotton (Gossypium hirsutum L.) genome
dc.contributor.authorZhang, J.
dc.contributor.authorCai, L.
dc.contributor.authorCheng, J.
dc.contributor.authorMao, H.
dc.contributor.authorFan, X.
dc.contributor.authorMeng, Z.
dc.contributor.authorChan, K.M.
dc.contributor.authorZhang, H.
dc.contributor.authorQi, J.
dc.contributor.authorJi, L.
dc.contributor.authorHong, Y.
dc.date.accessioned2014-12-12T07:35:13Z
dc.date.available2014-12-12T07:35:13Z
dc.date.issued2008-04
dc.identifier.citationZhang, J., Cai, L., Cheng, J., Mao, H., Fan, X., Meng, Z., Chan, K.M., Zhang, H., Qi, J., Ji, L., Hong, Y. (2008-04). Transgene integration and organization in Cotton (Gossypium hirsutum L.) genome. Transgenic Research 17 (2) : 293-306. ScholarBank@NUS Repository. https://doi.org/10.1007/s11248-007-9101-3
dc.identifier.issn09628819
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/116006
dc.description.abstractWhile genetically modified upland cotton (Gossypium hirsutum L.) varieties are ranked among the most successful genetically modified organisms (GMO), there is little knowledge on transgene integration in the cotton genome, partly because of the difficulty in obtaining large numbers of transgenic plants. In this study, we analyzed 139 independently derived T0 transgenic cotton plants transformed by Agrobacterium tumefaciens strain AGL1 carrying a binary plasmid pPZP-GFP. It was found by PCR that as many as 31% of the plants had integration of vector backbone sequences. Of the 110 plants with good genomic Southern blot results, 37% had integration of a single T-DNA, 24% had two T-DNA copies and 39% had three or more copies. Multiple copies of the T-DNA existed either as repeats in complex loci or unlinked loci. Our further analysis of two T1 populations showed that segregants with a single T-DNA and no vector sequence could be obtained from T0 plants having multiple T-DNA copies and vector sequence. Out of the 57 T-DNA/T-DNA junctions cloned from complex loci, 27 had canonical T-DNA tandem repeats, the rest (30) had deletions to T-DNAs or had inclusion of vector sequences. Overlapping micro-homology was present for most of the T-DNA/T-DNA junctions (38/57). Right border (RB) ends of the T-DNA were precise while most left border (LB) ends (64%) had truncations to internal border sequences. Sequencing of collinear vector integration outside LB in 33 plants gave evidence that collinear vector sequence was determined in agrobacterium culture. Among the 130 plants with characterized flanking sequences, 12% had the transgene integrated into coding sequences, 12% into repetitive sequences, 7% into rDNAs. Interestingly, 7% had the transgene integrated into chloroplast derived sequences. Nucleotide sequence comparison of target sites in cotton genome before and after T-DNA integration revealed overlapping microhomology between target sites and the T-DNA (8/8), deletions to cotton genome in most cases studied (7/8) and some also had filler sequences (3/8). This information on T-DNA integration in cotton will facilitate functional genomic studies and further crop improvement. © 2007 Springer Science+Business Media B.V.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1007/s11248-007-9101-3
dc.sourceScopus
dc.subjectAgrobacterium-mediated transformation
dc.subjectCotton (Gossypium hirsutum L.)
dc.subjectTransgene
dc.subjectTransgene repeats
dc.subjectTransgene/plant genome junctions
dc.subjectVector backbone integration
dc.typeArticle
dc.contributor.departmentINSTITUTE OF MOLECULAR AGROBIOLOGY
dc.description.doi10.1007/s11248-007-9101-3
dc.description.sourcetitleTransgenic Research
dc.description.volume17
dc.description.issue2
dc.description.page293-306
dc.description.codenTRSEE
dc.identifier.isiut000254353900012
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